Identifying the RP10 gene causing retinitis pigmentosa

描述（由申请人提供）：本项目的目的是鉴定，即“克隆”引起常染色体显性视网膜色素变性（adRP）的RP 10基因。在初步研究中，我们定位RP 10基因的人类染色体7 q31的连锁定位在4个大的，无关的adRP家庭。包含RP 10基因的区域，两侧是标记D 7S 2471和RP-STR 12，长度为3.3 mbp。通过结合公共（GenBank）和私人（Celera）人类基因组序列，我们已经组装了该区域99%的完整序列。同样，Celera数据库包含99%的同线小鼠序列。通过计算分析和实验室方法，我们已经确定了38个独立的，在该地区的表达序列，其中10个视网膜表达，因此，可能的疾病基因候选人。我们建议通过建立这些发现和资源来完成RP 10基因克隆的任务。 提出的目标是1）减少RP 10连锁区，2）完成该区域的物理和转录图谱，3）测试RP 10家族中的候选基因。目标1的方法包括增强的连锁分析和SNP单倍型测试，以完善断点;和生产单倍体细胞系，从患者，以促进测试。目标2的方法包括进一步的计算分析，比较小鼠和人类基因组;以及基于转录谱和功能对候选基因进行优先排序。目的3的方法包括使用基因敲除小鼠检测该区域的视网膜和光感受器表达基因;候选基因筛选;以及在无关adRP患者中确认阳性结果。这些研究将与南非国家生物信息学研究所和都柏林的Trinity学院的眼部遗传学单位合作进行。 RP 10基因的突变可能导致至少10%的adRP，这反过来又影响了成千上万的美国人。RP 10基因的鉴定将通过诊断和咨询为受影响的个体提供直接益处。此外，它是设计基因特异性疗法和鉴定修饰因子的必要的第一步。更广泛地说，导致视网膜色素变性的基因的鉴定揭示了视网膜中新的功能途径，因此克隆RP 10基因可能有助于更好地理解正常的视觉过程。

DESCRIPTION (provided by applicant): The purpose of this project is to identify, that is, "clone," the RP10 gene which causes autosomal dominant retinitis pigmentosa (adRP). In preliminary research we localized the RP10 gene to human chromosome 7q31 by linkage mapping in 4 large, unrelated adRP families. The region containing the RP10 gene, flanked by markers D7S2471 and RP-STR12, is 3.3 mbp in length. By combining the public (GenBank) and private (Celera) human genome sequences, we have assembled 99% of the complete sequence in this region. Likewise, the Celera database contains 99% of the syntenic mouse sequence. By computational analysis and laboratory methods we have identified 38 independent, expressed sequences in the region, 10 of which are retinal-expressed and, thus, possible disease-gene candidates. We propose to complete the task of cloning the RP10 gene by building on these findings and resources. Proposed aims are 1) to reduce the RP10 linkage region, 2) to complete the physical and transcriptional maps of the region and 3) to test candidate genes in RP10 families. Methods for Aim 1 include enhanced linkage analysis and SNP haplotype testing to refine breakpoints; and production of haploid cell lines from patients to facilitate testing. Methods for Aim 2 include further computational analyses, comparing mouse and human genomes; and prioritization of candidate genes based on transcript profiles and function. Methods for Aim 3 include detection of retinal and photoreceptor-expressed genes in the region using knockout mice; candidate gene screening; and confirmation of positive results in unrelated adRP patients. These studies will be done in collaboration with the South African National Bioinformatics lnstitute, and the Ocular Genetics Unit, Trinity College, Dublin. Mutations in the RP10 gene may cause at least 10% of adRP which, in turn, affects thousands of Americans. Identification of the RP10 gene will provide direct benefits to affected individuals through diagnosis and counseling. Also, it is a necessary first step in designing gene-specific therapies and identifying modifying factors. More broadly, identification of genes causing retinitis pigmentosa has revealed new functional pathways in the retina, thus cloning the RP10 gene will likely contribute to a better understanding of normal visual processes.